Proteomic Studies Of Dendrocalamopsis Beecheyana Var. Pubescens Under Different Salinity Stress

High salt concentration of the soil adverse to the most of the growth of vegetation.Dendrocalamopsis beecheyana var.pubescens as one of the tree species which has been planted successfully in the southeast coast which have strong adaptability locally.Thus,the research of salt-tolerant mechanism is especially important.Use the bamboo of biennial as experimental material and stress it by normal watering treatment and NaCl solution at the concentration of 0.4%and 0.6%.Build a system of dimensional electro-phoresis suitable for D.beecheyana var.pubescens to get the 2-DE map.The mass spectrum identification was used to determine the information function of differential protein,which provides a basis for a full understanding of the salt-tolerant mechanism of plants.The main results are as follows:(1)Compared with three methods of total protein extraction of Dendrocalamopsis beecheyana var.pubescens which include hydration operation mode of Strip,program arguments setting of isoelectric focusing and protein amount were optimized to establish an optimal Two-dimensional electrophoresis system suitable for Dendrocalamopsis beecheyana var.pubescens.Test optimizes the system of dimensional electropHoresis suitable for Dendrocalamopsis beecheyana var.pubescens.Bamboo proteins were mainly distributed in the scope of the pH 5-8.Using trichloroacetic acid-acetone method combined with phenol extraction method,placing the surface of strip up-side down,setting the isoelectric focusing of 65000V/H,using 24 hours passive hydration mode,and 50?L amount of protein are benefit to obtain optimal effect of gel electrophoresis.(2)Using the optimal Two-dimensional electrophoresis system suitable for Dendrocalamopsis beecheyana var.pubescens to acquire about 600 proteins in statistically significant differences detected by PDQuest8.0.1.Under the stress of 0.4%salt concentration,there are 34 differentially expressed proteins(12 increased and 22 decreased).Under the stress of 0.6%salt concentration,there are 40 differentially expressed proteins(13 increased and 27 decreased).(3)Eighteen differential proteins were selected for mass spect rometry identification,fifteen of them were identified successfully.Among them,under the stress of 0.4%and 0,6%salt concentration,the protein expression of Oxygen enhancer protein 1?Oxygen enhancer protein 3?phosphoglycerate kinase and ribulose-1,5-bisphosphate carboxylase increased when the concentration was increased.The protein expression of Glutamine synthelase and Ascorbic acid peroxidase 2 significantly increased.The protein expression of ATP synthase beta subunit and ATP synthase significantly down regulate.The protein expression of ATP synthase alpha subunit rise in the first stage,and then decrease.(4)By the GO(Gene Ontology)comments and KEGG analysis in 15 different proteins,most of the differences in protein in cells which mainly involved in cellular and metabolic process.Its main function are Ion combined with function?Transfer activity function and Catalytic activity function.Its main involved photosynthesis(4 protein spots).Oxidative phosphorylation(3 protein spots)and Carbon metabolism(5 protein spots).They are divided into 5 classes which involves photosynthesis related proteins(20%)?Protein involved in energy metabolism(20%)?Carbon metabolism related proteins(34%)?Cell defense protein related proteins(13%)?others and unknown protein(13%).(5)The test found that Ribulose-l,5-bisphosphate carboxylaseGlutamine synthelase(Cell defense related proteins)and Ascorbic.acid peroxidase 2(Tolerant protein)can express abundant in response to salt stress.They have important effect in salt-tolerance mechanism of Dendrocalamopsis beecheyana var.pubescens.